Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
HRI CRISPR Activation Plasmid (h): sc-403442-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HRI CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- HRI CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by HRI CRISPR Activation Plasmid (h) and HRI CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the EIF2AK1 transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HRI Antibody (D-12): sc-365239
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HRI CRISPR Activation Plasmid (h) | sc-403442-ACT | 20 µg | $397.00 |
EIF2AK1 encodes heme-regulated inhibitor (HRI), a stress-responsive eIF2α kinase that couples heme availability and oxidative or proteotoxic stress to control of translation initiation. Upon activation, HRI phosphorylates EIF2S1/eIF2α to reduce global protein synthesis while promoting integrated stress response programs that support redox balance and proteostasis. This pathway intersects with heme metabolism, mitochondrial function, and cytosolic quality control, influencing erythroid maturation and broader cellular adaptation to iron and heme limitation. Dysregulated HRI–eIF2α signaling has been associated with altered stress tolerance and inflammatory signaling relevant to anemia-related biology and neurodegeneration-linked proteostasis defects.
HRI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF2AK1 expression without altering the underlying DNA sequence.
HRI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF2AK1 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.
Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the EIF2AK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HRI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF2AK1 locus and enabling the study of HRI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HRI pathway restoration in tumor cells with silenced or reduced EIF2AK1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.